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Objective:To investigate the expression of high mobility group protein 1 (HMGB1) and interleukin-17 (IL-17) in peripheral blood and membrane tissues of pregnant women with premature rupture of membranes (PROM) and its relationship with intrauterine infection.Methods:Seventy-four pregnant women with PROM from January 2019 to June 2021 were selected as the study group, and 58 healthy pregnant women at the corresponding period were selected as the healthy control group. The levels of HMGB1 and IL-17 in peripheral blood and membrane tissues and serum CD 8+ were compared between the two groups. The pregnant women with PROM were divided into the chorioamnionitis group, subclinical chorioamnionitis group and normal group according to their intrauterine infection, the expression levels of HMGB1 and IL-17 in peripheral blood and membrane tissues of patients with different infection degrees were compared, and the correlation with the severity of intrauterine infection were analyzed. Results:The levels of peripheral blood HMGB1, membrane tissues HMGB1, peripheral blood IL-17, membrane tissues IL-17 and serum CD 8+ in the study group were higher than those in the control group: (28.34 ± 5.16) μg/L vs. (22.51 ± 4.09) μg/L, 0.79 ± 0.12 vs. 0.34 ± 0.05, (13.05 ± 2.57) ng/L vs. (8.16 ± 1.38) ng/L, 0.37 ± 0.06 vs. 0.12 ± 0.02, 0.386 ± 0.052 vs. 0.252 ± 0.044, there were statistical differences ( P<0.05). The levels of HMGB1 and IL-17 in peripheral blood and membrane tissues and serum CD 8+ were increased with the severity of severity of intrauterine infection ( P<0.05). The results of Spearman correlation analysis showed that the level of peripheral blood HMGB1, membrane tissues HMGB1 and IL-17 had positively correlated with the severity of intrauterine infection ( r = 0.336, 0.316, 0.311, P<0.05). The results of receiver operating characteristic curve analysis showed that combined detection of HMGB1 and IL-17 levels in peripheral blood and membrane tissues and serum CD 8+ levels in evaluating the severity of intrauterine infection had higher area under the curve than that of each index alone ( P<0.05). Conclusions:Pregnant women with PROM have abnormal HMGB1 and IL-17 levels in peripheral blood and membrane tissues, and HMGB1 levels in peripheral blood and mRNA expressions of HMGB1 and IL-17 in membrane tissues are positively correlated with the severity of intrauterine infection, which has evaluation value for the severity of the disease.
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Objective:To investigate the clinical significance of the expression of oxidized α1-antitrypsin (ox-AAT) and neutrophil elastase (NE) in the peripheral blood and fetal membrane tissues of pregnant women with premature rupture of membranes (PROM).Methods:The clinical data of 95 cases of PROM pregnant women admitted to Binhai County People′s Hospital from April 2019 to April 2020 were analyzed. According to combination of histological chorioamnionitis (HCA), they were divided into PROM combined with HCA group ( 31 patients) and PROM without HCA group (64 patients). Besides, 50 normal pregnant women were collected during the same period as a healthy control group. The expression levels of ox-AAT and NE in the peripheral blood and fetal membrane tissues of the three groups were compared and analyzed.Results:The levels of peripheral blood ox-AAT and NE in the PROM combined with HCA group were higher than those in PROM without HCA group and healthy control group: (2.34 ± 0.02) ng/L vs. (1.50 ± 0.12), (0.32 ± 0.04) ng/L; (0.48 ± 0.08) ng/L vs. (0.13 ± 0.06), (0.11 ± 0.05) ng/L;the level of peripheral blood ox-AAT in PROM without HCA group was higher than that in healthy control group: (1.50 ± 0.12) ng/L vs. (0.32 ± 0.04) ng/L, and the differences were statistically significant ( P<0.05). The levels of fetal membrane tissues ox-AAT and NE in the PROM combined with HCA group were higher than those in PROM without HCA group and healthy control group: (0.031 ± 0.005) ng/L vs. (0.015 ± 0.002), (0.009 ± 0.003) ng/L; (0.020 ± 0.002) ng/L vs. (0.003 ± 0.001), (0.002 ± 0.001) ng/L; the level of fetal membrane tissues ox-AAT in PROM without HCA group was higher than that in the healthy control group: (0.015 ± 0.002) ng/L vs. (0.009 ± 0.003) ng/L, and the differences were statistically significant ( P<0.05). There was a positive correlation between ox-AAT and NE in peripheral blood and fetal membrane tissues ( r = 0.879, 0.875, P<0.05). The incidence of placental abruption in the PROM combined with HCA group and PROM without HCA group was higher than that in the healthy control group: 32.26%(10/31), 20.31%(13/64) vs. 4.00%(2/50), the incidence of neonatal pneumonia in the PROM combined with HCA group was higher than that in the PROM without HCA group and healthy control group: 25.81%(8/31) vs. 9.38%(6/64), 2.00%(1/50), and the differences were statistically significant ( P<0.05). Conclusions:The level of ox-AAT is overexpressed in peripheral blood and fetal membrane tissues of pregnant women with PROM, the level of NE is overexpressed in peripheral blood and fetal membrane tissues of PROM combined with HCA, and the increase of ox-AAT and NE expression is closely related to adverse perinatal outcomes.
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Objective:To investigate the effect of CD 8+ CD 25+ FoxP3 + regulatory T cell (Treg) expression levels in peripheral blood of pregnant women with premature rupture of fetal membranes(PROM) on immune function of helper T cells (Th) 1/Th2. Methods:Thirty cases of pregnant women with PROM (PROM group), 30 cases of normal pregnant women (normal pregnancy group) and 30 cases of normal non-pregnant women (non-pregnancy group) who treated in Binhai County People′s Hospital from September 2019 to May 2020 were collected. Peripheral blood of each group was collected and the proportion of CD 8+ CD 25+ FoxP3 + Treg was determined by flow cytometry. Peripheral blood mononuclear cells (PBMCs) were extracted and FoxP3 mRNA was determined by polymerase chain reaction (PCR). The levels of Th1-related cytokines interferon-γ (IFN-γ), interleukin (IL)-2, and Th2-related cytokines IL-10 and IL-4 were measured by Luminex liquid phase microarray. The effects of CD 8+ CD 25+ FoxP3 + Tregexpression on Th1/Th2 balance were analyzed. Results:The proportion of CD 8+ CD 25+ FoxP3 + Tregand the expression of FoxP3 mRNA in PROM groupand normal pregnancy group were lower than those in non-pregnancy group: (0.15 ± 0.03) %, (0.35 ± 0.09) % vs. (0.47 ± 0.11) %; 0.89 ± 0.11, 3.15 ± 0.67 vs. 3.75 ± 0.23 , the proportion of CD 8+ CD 25+ FoxP3 + Treg and the expression of FoxP3 mRNA in PROM groupwere lower than those in the normal pregnancy group , and the differences were statistically significant ( P<0.05). The levels of Th1-related cytokines IFN-γ and IL-2 in PROM group and normal pregnancy group were higher than those in non-pregnancy group, the level of Th2-related cytokines IL-4 was lower than that in non-pregnancy group , the levels of IFN-γ and IL-2 in PROM group were higher than those in normal pregnancy group, the level of IL-4 was lower than that in normal pregnancy group , and the differences were statistically significant ( P<0.05). In PROM group, the proportion of CD 8+ CD 25+ FoxP3 + Treg and the expression of FoxP3 mRNA in peripheral blood were negatively correlated with Th1-related cytokines IFN-γ ( r = - 0.413, -0.451, P<0.05) and IL-22 ( r = -0.645, -0.535, P<0.05), and were positively correlated with Th2-related cytokines IL-4 ( r = 0.558, 0.469, P<0.05). Conclusions:The proportion of CD 8+ CD 25+ FoxP3 + Treg in peripheral blood of pregnant women with PROM is lower, and the expression level of related FoxP3 mRNA is lower, which all affecte the Th1/Th2 immune balance and cause Th1 immune drift, which may be the related immune mechanism of PROM.
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AIM: To investigate the effects of propofol on the expression of tissue-type plasminogen activator (tPA) and matrix metalloproteinase 9 (MMP9) in the hippocampus and the cognitive function in neonatal rats.METHODS: The 7-day-old rats were randomly divided into 3 groups: the rats in control (CON) group were intraperitoneally injected with normal saline for 7 d;the rats in single dose of propofol anesthesia (SP) group were intraperitoneally injected with normal saline for 6 d and with propofol on the 7th day;the rats in repeated dose of propofol anesthesia (RP) group were intraperitoneally injected with propofol for 7 d.Blood glucose and blood gas analysis were tested in 6 rats of each group.The rats were randomly selected from each group to isolate the hippocampal tissues at 2 h, 24 h, 48 h, 72 h and 30 d after the last injection.The spatial learning and memory functions of the other rats aged 25 d were determined by Morris water maze.The morphological changes of the hippocampus were observed by HE staining and Nissl's staining.The expression of tPA and MMP9 at mRNA and protein levels was determined by RT-PCR and Western blot.RESULTS: Compared with group CON, the protein expression of tPA and MMP9 in RP group was significantly decreased at each time point, while no significant decrease was observed in SP group except at the time point of 24 h.Compared with CON group, the mRNA expression of tPA and MMP9 was down-regulated obviously in RP group, which was not significantly down-regulated in SP group.From the 3rd training day of Morris water maze beginning, the escape latency was prolonged, and the space exploration time and the number of crossing the original platform location were reduced in RP group compared with CON group and SP group, while no significant difference was observed between CON group and SP group.Compared with CON group, the number of nerve cells reduced and nerve cells arranged in disorder in the hippocampus in RP group.Moreover, the number of Nissl body decreased significantly and finally developed into neuronal degeneration and necrosis in RP group, and no significant difference between SP group and CON group was observed.CONCLUSION: Repeated dose of propofol anesthesia leads to long-term cognitive dysfunction in neonatal rats, which may be related to the down-regulation of tPA and MMP9 expression and destruction of normal morphology and function of neurons in hippocampus, whereas single dose of propofol anesthesia has no such effects.
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ObjectiveTo explore the role of peripheral blood intermediate monocytes in the pathogenesis of preeclampsia.MethodsFifty-two patients with established preeclampsia in Binhai County People's Hospital from October 2014 to October 2015, 42 healthy pregnant women and 42 healthy non-pregnant women were enrolled in this study. The percentage of intermediate monocyte subsets, ratio of positive cells and mean fluorescence intensity (MFI) of Toll-like receptor (TLR) 2, TLR4, CD64, and triggering receptor expressed on myeloid cell-1(TREM-1), and MFI of intracellular tumor necrosis factor-α (TNF-α) and interleukin (IL)-6 were evaluated by flow cytometry. The concentrations of IL-6, IL-8, IL-1β, IL-12P70 and TNF-α in serum were analyzed using Luminex liquid phase chip technology. Independent two samplest-test, analysis of variance, Mann-WhitneyU test, Kruskal-Wallis test and Pearson correlation analysis were used for statistical analysis.ResultsPercentage of intermediate monocytes was higher in preeclampsia patients [10.4%(5.3%-19.9%)]than in healthy pregnant women [6.6%(4.9%-7.8%)], and both were higher than in non-pregnant women [3.8%(2.4%-5.0)%](allP<0.05). The ratio of TLR4 and CD64 positive intermediate monocytes [(60.1±12.5)%vs (24.9±8.8)%; (85.3±5.4)% vs (67.4±7.5)%](t were 15.416 and 13.437, bothP<0.05), and MFI of TLR4 (50.3±10.2 vs 26.8±8.6), TREM-1(35.6±4.1 vs 28.6±4.7) and CD64 (39.8±5.2 vs 28.9±4.8) (t were 11.898, 7.707 and 10.454, allP<0.05) were higher in preeclampsia patients than in healthy pregnant women. MFI of intracellular IL-6 (32.3±4.7 vs 28.6±3.5) and TNF-α (44.6±6.3 vs 36.7±8.3) in intermediate monocytes of preeclampsia patients was also significantly higher than that of healthy pregnant women (t were 4.239 and 5.245, bothP<0.05). Serum concentrations of IL-6, IL-8 and TNF-α were higher in preeclamptic patients than in healthy pregnant women and non-pregnant women (allP<0.05). Furthermore, a positive correlation was found between the percentage of intermediate monocytes and the serum levels of IL-6 and TNF-α in preeclamptic patients (r were 0.397 and 0.347, bothP<0.05).ConclusionsMonocyte subpopulations from preeclamptic patients are abnormally skewed toward intermediate monocytes which have high expressions of TLR4, TREM-1 and CD64, and secret more proinflammatory cytokines such as IL-6 and TNF-α. Therefore, intermediate monocytes are specifically altered in preeclamptic patients and may play a role in the pathophysiology of preeclampsia.
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Aim To investigate the role of matrix metalloproteinase-9 down-regulation in the learning and memory dysfunction induced by propofol treatment in rats.Methods 7-day-old SD rats were randomly divided into three groups(n=18):control group(NS group) and repeated doses of propofol group(RP group) was intraperitoneally injected with normal saline and propofol respectively for consecutive seven days, single dose of propofol group(SP group) were intraperitoneally injected with normal saline first for consecutive six days, and then injected with propofol on 7th day.The blood gas and glucose levels were monitored of six rats randomly selected from each group.Morris water maze was conducted to test the learning and memory functions of the remaining rats.The expression of MMP-9, BDNF and caspase-3 was detected by Western blot, and the hippocampal neuron apoptosis was determinated by TUNEL staining.Results Compared with NS group and SP group, the escape latency in RP group was prolonged significantly, exploration time and the number of crossing the platform in RP group were markedly decreased(P0.05).Conclusions Repeated exposure to propofol can lead to a decline in long-term learning and memory functions in neonatal rats, which may be related to the down-regulation of MMP-9 expression, proBDNF and mBDNF conversion disorder in hippocampus and the apoptosis of hippocampal neurons.However, single exposure to propofol has no significant effect.
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Aim To observe the improvement effects of parecoxib on ventricular remodeling after acute myocardial infarction in rats and its influence on PI3K/Akt signaling pathway.Methods Forty male Sprague-Dawley rats were randomly divided into five groups(n=8):sham operation group (group S), ventricular remodeling model group(group R), low dose of parecoxib group(group P1), middle dose of parecoxib group(group P2), and high dose of parecoxib group(group P3).A myocardial infarction model was established by ligating the left anterior descending branch(LAD) of coronary artery in group R, group P1, group P2 and group P3.One day after the operation,the rats were given intraperitoneal injection of 4,8,12 mg·kg-1 parecoxibin Group P1, group P2 and group P3,respectively, for two weeks.The same volume saline was given in group S and group R.Four weeks later, LVSP, LVEDP,+dp/dtmax,-dp/dtmax were monitored.The hearts were harvested to calculate left ventricular hypertrophy index.The pathological change of heart was examined with an optical microscope.The expressions of atrial natriuretic peptide(ANP) mRNA and brain natriuretic peptide(BNP) mRNA were detected by RT-PCR.The expression of PI3K,Akt,P-Akt and caspase-3 was detected by Western blot.Results Compared with group S, the cardiac function was decreased, the left ventricular hypertrophy index, the expression levels of ANP,BNP mRNA, caspase-3 were increased, and the expression levels of PI3K, P-Akt were reduced in group R(all P<0.05).Compared with group R, the cardiac function was ameliorated, the left ventricular hypertrophy index were reduced in group P2 and group P3(all P<0.05).The expression levels of ANP,BNP mRNA, Caspase-3 were decreased, and the expression levels of PI3K and P-Akt were increased in group P1,group P2 and group P3(all P<0.05).Conclusions Middle and high doses of parecoxib can mitigate the process of ventricular remodeling after myocardial infarctionand improve the myocardial function, and its underlying mechanism may be related to activating PI3K/Akt signaling pathway.
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Objective:To explore the rule of CD8+CD25+FoxP3+ regulator T cells(Treg) in the pathogenesis of pre-eclampsia (PE) in peripheral blood.Methods:This study included 24 gestational-age-matched healthy pregnant women and 46 pregnant women diagnosed with mild PE (MPE,n=24) or severe PE (SPE,n=22) during the third trimester of gestation.An 3 ml sample of peripheral blood was drawn from each subject and anti-coagulated with heparin sodiurm The percentage of CD8 + CD25 + FoxP3 + Treg cells was detected by flow cytometry.The cytokines IL-6,IL-17A,IL-10,IL-1,IL-33 and TGF-β31 were detected using Luminex200.Peripheral blood mononuclear cells (PBMCs) were isolated frum healthy controls and treated with IL-33,the percentages of CD8+ CD25+ FoxP3 + Treg cell were measured by flow cytometric detection.Results:Compared to that of healthy pregnant controls [0.48(0.21-0.96)％],MPE patients [0.32(0.19-0.63)％] and SPE patients [0.13(0.02-0.41)％] had lower percentages of CD8+CD25+FoxP3+Treg cells (P＜0.05).Compared to HP controls,higher levels of IL-6 and IL-17A were found in MPE and SPE patients(P＜0.05) and even higher in SPE patients.The levels of IL-10,IL-1[β and IL-33 were similar in all three groups (P＞0.05).Compared to HP contruls,the levels of TGF-[β1 was significantly increased in SPE and MPE patients(P＜0.05),but no significant differences were found between these two groups (P ＞ 0.05).The percentage of CD8+ CD25+ FoxP3+ Treg cells showed a negative correlation with the serum concentrations of IL-17A (r =-0.338,P =0.338),and a positive correlation with the serum concentrations of IL-33 (r =0.548,P =0.548).After PBMCs were treated with IL-33 for five days,the percentages of CD8+CD25+FoxP3+ Treg were significantly higher than those of the contruls (P＜0.05).Conclusion:These findings suggested that the reduced CD8+ CD25 + FoxP3 + Treg cells may play a role in the pathogenesis of ore-eclamosia.
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Objective To compare the differences between laparoscopic-assisted and open resection of rectal carcinoma in respect of peri-operative levels of C-reactive protein (CRP), rest energy expenditure (REE), and visceral proteins. Methods According to patients' choice of operation, either laparoscopic-assisted (n=20, Laparoscopic Group) or open (n=25, Open Group) resection of rectal carcinoma was performed. The levels of CRP and visceral proteins-including albumin (ALB), prealbumin (PRE), transferrin (TRF), and retinal-binding protein (RbP)-were assayed preoperatively and on the 1st, 2nd, and 3rd day postoperatively. The levels of REE were also measured by indirect calorimetry in the morning. Results Compared with the preoperative period, the CRP levels in both groups were significantly increased on the 1st, 2nd, and 3rd day (P0.05). The levels of PRE in the Laparoscopic Group were significantly higher than those in the Open Group on the 2nd postoperative day (P